Journal: bioRxiv

Article Title: AGP-Ca 2+ binding is essential for pollen development and pollen tube growth in Arabidopsis thaliana

doi: 10.64898/2026.01.27.701722

Figure Lengend Snippet: a , Alexander staining of anthers at flower stage 12 and pollen grains (PGs) of anthers at flower stage 13 of wild-type (WT), glcat14a/b/d-1 , and glcat14a/b/d-2 plants. Viable PGs are stained in red-magenta while aborted PGs are stained in blue-green (black arrows). b , Viable and aborted PGs percentage of WT and glcat14a/b/d mutants. Data are presented as mean + SEM ( n = 3 plants, 500 PGs per plant). Mean values are shown within bars. Statistical significance was determined by two-way ANOVA followed by Dunnett’s test. Asterisks represent statistically significant differences from WT (****, p <0.0001). c , Aniline blue staining of anthers at flower stage 13 of WT and glcat14a/b/d plants. d , DAPI staining of PGs of anthers at flower stage 13 of WT and glcat14a/b/d mutants. Sperm cell (white arrows) and vegetative (white asterisks) nuclei are shown in the DAPI channel, alongside the corresponding brightfield channel indicating PG boundaries. e , Anther sections at flower stages 9–12, showing pollen mother cell, tetrad, uninucleate microspore, bicellular, and tricellular PG stages of WT and glcat14a/b/d plants. Abbreviations: cp, collapsed pollen grain; dm, degenerated microspore; en, endothecium; ep, epidermis; ml, middle layer; ms, microspore; pg, pollen grain; PMC, pollen mother cell; t, tapetum; td, tetrad. Images were acquired using a Zeiss Axio Imager.A2 microscope with DIC optics ( a , e ), Leica DMLB epifluorescence microscope ( c ), and Leica TCS SP5 II confocal system ( d ). Scale bars, 50 μm.

Article Snippet: DIC images were acquired using a Zeiss Axio Imager.A2 upright microscope with DIC optics, equipped with a Zeiss AxioCam MRc 3 camera and a 60N-C 2/3 0.63× camera adapter.

Techniques: Staining, Microscopy

Journal: bioRxiv

Article Title: AGP-Ca 2+ binding is essential for pollen development and pollen tube growth in Arabidopsis thaliana

doi: 10.64898/2026.01.27.701722

Figure Lengend Snippet: a , Autofluorescence, calcofluor white, decolourised aniline blue solution (DABS), and GFP–CBM3 staining of anther sections at flower stage 12 of wild-type (WT), glcat14a/b/d-1 , and glcat14a/b/d-2 plants. b , Transmission electron microscopy of pollen grains (PGs) of WT and glcat14a/b/d mutans. c , In vitro pollen germination of PGs of WT and glcat14a/b/d plants. Abbreviations: ba, bacula; cp, collapsed pollen grain; en, endothecium; ep, epidermis; ex, exine; in, intine; ne, nexine; pc, pollen coat; pg, pollen grain; pm, plasma membrane; pt, pollen tube; te, tectum. Images were acquired using a Leica TCS SP8 confocal system ( a ), JEOL JEM 1400 transmission electron microscope ( b ), and Zeiss Axio Imager.A2 microscope with DIC optics ( c ). Scale bars, 1 μm ( b ), 50 μm ( a ), and 100 μm ( c ). d , e , Pollen germination percentage ( d ) and pollen tube (PT) length ( e ) of WT and glcat14a/b/d mutants. Data are presented as mean + SEM [ n = 3 plants, 150 PGs per plant for ( d ) and 50 PTs per plant for ( e )]. Biological replicates are shown as black dots. Mean values are shown within bars. Statistical significance was determined by one-way ANOVA followed by Dunnett’s test. Asterisks represent statistically significant differences from WT (**, p <0.0021; ****, p <0.0001).

Article Snippet: DIC images were acquired using a Zeiss Axio Imager.A2 upright microscope with DIC optics, equipped with a Zeiss AxioCam MRc 3 camera and a 60N-C 2/3 0.63× camera adapter.

Techniques: Staining, Transmission Assay, Electron Microscopy, In Vitro, Clinical Proteomics, Membrane, Microscopy

Journal: bioRxiv

Article Title: AGP-Ca 2+ binding is essential for pollen development and pollen tube growth in Arabidopsis thaliana

doi: 10.64898/2026.01.27.701722

Figure Lengend Snippet: a , Alexander staining of anthers at flower stage 12 and pollen grains (PGs) of anthers at flower stage 13 of wild-type (WT), glcat14a/b/d-1 , and glcat14a/b/d-2 grown with basal nutrient solution (BNS) and low calcium nutrient solution (LCNS). Viable PGs are stained in red-magenta while aborted PGs are stained in blue-green (black arrows). b , Viable and aborted PGs percentage of WT and glcat14a/b/d mutants grown with BNS and LCNS. Data are presented as mean + SEM ( n = 3 plants, 500 PGs per plant). Mean values are shown within bars. Statistical significance was determined by two-way ANOVA followed by Tukey’s test. Asterisks represent statistically significant differences from WT grown in BNS or LCNS, unless otherwise specified. Magenta asterisks indicate significance for viable pollen, and green asterisks indicate significance for aborted pollen (*, p <0.0332; ****, p <0.0001). c , DAPI staining of PGs of anthers at flower stage 13 of WT and glcat14a/b/d mutants. Sperm cell (white arrows) and vegetative (white asterisks) nuclei are shown in the DAPI channel, alongside the corresponding brightfield channel indicating PG boundaries. d , Anther sections at flower stage 12 showing tricellular PG stages of WT and glcat14a/b/d plants. Abbreviations: cp, collapsed pollen grain; dt, degenerated tapetum; en, endothecium; ep, epidermis; pg, pollen grain. Images were acquired using a Zeiss Axio Imager.A2 microscope with DIC optics ( a , d ) and Leica TCS SP5 II confocal system ( c ). Scale bars, 50 μm.

Article Snippet: DIC images were acquired using a Zeiss Axio Imager.A2 upright microscope with DIC optics, equipped with a Zeiss AxioCam MRc 3 camera and a 60N-C 2/3 0.63× camera adapter.

Techniques: Staining, Microscopy

Journal: bioRxiv

Article Title: AGP-Ca 2+ binding is essential for pollen development and pollen tube growth in Arabidopsis thaliana

doi: 10.64898/2026.01.27.701722

Figure Lengend Snippet: a , In vitro pollen germination of PGs of wild-type (WT), glcat14a/b/d-1 , and glcat14a/b/d-2 grown with BNS and LCNS. Abbreviations: cp, collapsed pollen grain; pg, pollen grain; pt, pollen tube. Images were acquired using a Zeiss Axio Imager.A2 microscope with DIC optics Scale bars, 100 μm. b , c , Pollen germination percentage ( b ) and pollen tube (PT) length ( c ) of WT and glcat14a/b/d mutants 2 grown with BNS and LCNS. Data are presented as mean + SEM [ n = 3 plants, 150 PGs per plant for ( b ) and 50 PTs per plant for ( c) ]. Biological replicates are shown as black dots. Mean values are shown within bars. Statistical significance was determined by two-way ANOVA followed by Tukey’s test. Asterisks represent statistically significant differences from WT (*, p <0.0332; **, p <0.0021; ***, p <0.0002 ****, p <0.0001).

Article Snippet: DIC images were acquired using a Zeiss Axio Imager.A2 upright microscope with DIC optics, equipped with a Zeiss AxioCam MRc 3 camera and a 60N-C 2/3 0.63× camera adapter.

Techniques: In Vitro, Microscopy